Journal: Blood Coagulation & Fibrinolysis

Article Title: Characterization of antibody clones that bind exclusively to insoluble fibrin

doi: 10.1097/MBC.0000000000001171

Figure Lengend Snippet: Fibrin deposition stained with anti-fibrin antibodies in mouse pancreatic tissue samples. (a) Frozen tissue samples were incubated with 1 μg/ml of HRP-conjugated anti-fibrin antibodies. (b) FFPE tissue samples were incubated with 10 μg/ml of HRP-conjugated anti-fibrin antibodies. The arrows indicate islets of Langerhans. HE staining is shown in the leftmost column. Scale bar, 100 μm. HRP, horseradish peroxidase.

Article Snippet: A frozen human PDAC tissue section (OriGene Technologies, Maryland, USA) and a normal human pancreatic tissue section (BioChain, California, USA) were purchased commercially.

Techniques: Staining, Incubation

Journal: Blood Coagulation & Fibrinolysis

Article Title: Characterization of antibody clones that bind exclusively to insoluble fibrin

doi: 10.1097/MBC.0000000000001171

Figure Lengend Snippet: Fibrin deposition stained with anti-fibrin antibodies in human pancreatic tissue samples. (a) Frozen tissue samples were incubated with 1 μg/ml of HRP-conjugated anti-fibrin antibodies. (b) FFPE tissue samples were incubated with 10 μg/ml of HRP-conjugated anti-fibrin antibodies. The arrow indicates islets of Langerhans. HE staining is shown in the leftmost column. Scale bar, 100 μm. HRP, horseradish peroxidase.

Article Snippet: A frozen human PDAC tissue section (OriGene Technologies, Maryland, USA) and a normal human pancreatic tissue section (BioChain, California, USA) were purchased commercially.

Techniques: Staining, Incubation

Journal: Blood Coagulation & Fibrinolysis

Article Title: Characterization of antibody clones that bind exclusively to insoluble fibrin

doi: 10.1097/MBC.0000000000001171

Figure Lengend Snippet: Improvement of human pancreatic tissue staining with HRP-conjugated 99 and 1101 clones. Frozen tissue samples were incubated with 10 μg/ml of HRP-conjugated 99 or 1101 antibody. Scale bar, 100 μm. HRP, horseradish peroxidase.

Article Snippet: A frozen human PDAC tissue section (OriGene Technologies, Maryland, USA) and a normal human pancreatic tissue section (BioChain, California, USA) were purchased commercially.

Techniques: Staining, Clone Assay, Incubation

Journal: PLoS ONE

Article Title: Roles of DPY30 in the Proliferation and Motility of Gastric Cancer Cells

doi: 10.1371/journal.pone.0131863

Figure Lengend Snippet: (A-D) Immunohistochemical staining demonstrated the overexpression of DPY30 in gastric cancer tissues. Notably its overexpression was obviously greater in invading cancer cells (B-D) than in normal gastric mucosa (A). (E) The mRNA level of DPY30 in gastric cancer cells (SNU1, SNU16, SNU216, SNU620, SNU638 and NCI-N87) and normal gastric epithelial cell (HFE145) was determined by real-time PCR using specific primers for DPY30. GAPDH was used to normalize data. Values shown are the means ± SDs of the three independent experiments performed in triplicate. *, p < 0.01 (Student’s t test, versus HFE145). (F) The expression of DPY30 in gastric cancer tissues was examined by real-time PCR using specific primers for DPY30. GAPDH was used to normalize data. Values are the means ± SDs of three independent experiments performed in triplicate. *, p < 0.01; **, p < 0.05 (Student’s t test, versus normal).

Article Snippet: Immunohistochemical staining with rabbit anti-human DPY30 polyclonal antibody (Sigma-Aldrich) was performed on a gastric cancer tissue array (n = 59) purchased from SUPER BIO CHIPS (Seoul) or on 4-μm sections of paraffin-embedded specimens.

Techniques: Immunohistochemical staining, Staining, Over Expression, Real-time Polymerase Chain Reaction, Expressing

Journal: Nature Communications

Article Title: Palmitoylation-driven PHF2 ubiquitination remodels lipid metabolism through the SREBP1c axis in hepatocellular carcinoma

doi: 10.1038/s41467-023-42170-0

Figure Lengend Snippet: a Levels of palmitoyl-coenzyme A (palmitoyl-CoA) were quantified using liquid chromatography-mass spectrometry (LC-MS) in HepG2 cells treated by palmitic acid (PA) treatment for 24 h. Mean ± SD ( n = 3 independent samples); * P < 0.05. Statistical analyses were based on a two-tailed unpaired t test. The exact p-values are presented in Supplementary Data . b Hepatocellular carcinoma (HCC) cells were pre-treated with 2-bromopalmitate (2-BP, 50 µM) or dimethyl sulfoxide (DMSO) for 24 h and incubated with PA for 24 h, followed by MG132 (10 µM) for 8 h. Immunoprecipitation was performed using an anti-PHF2 antibody. The precipitated proteins were subjected to an acyl-biotin exchange (ABE) assay using hydroxylamine (HAM) treatment to remove PA from palmitoylated cysteine residues. Free cysteines were labeled with BMCC-Biotin. Finally, palmitoylated proteins were detected using streptavidin-HRP. n = 3 independent experiments. IP: immunoprecipitation; Palm-PHF2: palmitoylated PHF2. c The palmitoylation site within PHF2 was identified using LC-MS analysis. The y and b fragments detected are as indicated in the sequence. The palmitoylation of cysteine 23 residue (C23) in PHF2 was identified based on the shift of peptide peaks. d After transfection with wild-type (WT) PHF2 or C23A-mutated PHF2 plasmids, HCC cells lysates were subjected to western blotting. n = 3 independent experiments. e HCC cells were transfected with Flag-PHF2-WT or Flag-PHF2-C23A mutant. The expressed proteins were immunoprecipitated with Flag affinity beads and subjected to ABE assay. The palmitoylated proteins were detected using western blotting. n = 3 independent experiments. Source data are provided as a file.

Article Snippet: Statistical analyses were based on a two-tailed unpaired t-test. c – e Analysis of a human HCC tissue array containing 38 pairs of clinical HCC with corresponding nine normal tissues (SuperBioChips Lab, Seoul, South Korea). c (Left) Representative images of HCC tissues immunostained with the indicated antibodies.

Techniques: Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Two Tailed Test, Incubation, Immunoprecipitation, Labeling, Sequencing, Residue, Transfection, Western Blot, Mutagenesis

Journal: Nature Communications

Article Title: Palmitoylation-driven PHF2 ubiquitination remodels lipid metabolism through the SREBP1c axis in hepatocellular carcinoma

doi: 10.1038/s41467-023-42170-0

Figure Lengend Snippet: a The profile of gene expressions of liver tissues obtained from the NCBI GEO dataset (GSE54238). A heatmap of gene expressions was displayed based on ZDHHC23 expression values. The color scale bar represents relative expression values ranging from low (blue) to high (yellow). b The mRNA levels were measured in HCC tissues and adjacent normal liver tissues from 30 HCC patients. Comparisons were performed by the unpaired t-test; mean ± SD ( n = 30 independent liver tissues). Statistical analyses were based on a two-tailed unpaired t-test. c – e Analysis of a human HCC tissue array containing 38 pairs of clinical HCC with corresponding nine normal tissues (SuperBioChips Lab, Seoul, South Korea). c (Left) Representative images of HCC tissues immunostained with the indicated antibodies. Scale bar = 60 µm. (Right) The staining scores were calculated based on immune-positive cell numbers; mean ± SD; P values were calculated by a two-tailed unpaired t-test. HPF a high-power field; N adjacent normal tissues; T HCC tissues. d A simple linear regression analysis examined correlations between proteins in HCC tissues. r 2 , a measure of goodness-of-fit, is calculated by comparing the sum-of-squares from the regression line with the sum-of-squares. e Kaplan–Meier analysis of the survival of HCC patients. Blue and red lines represent the indicated proteins’ low- and high-expression groups, respectively. P values were calculated using the log-rank test.

Article Snippet: Statistical analyses were based on a two-tailed unpaired t-test. c – e Analysis of a human HCC tissue array containing 38 pairs of clinical HCC with corresponding nine normal tissues (SuperBioChips Lab, Seoul, South Korea). c (Left) Representative images of HCC tissues immunostained with the indicated antibodies.

Techniques: Expressing, Two Tailed Test, Staining